胡卢巴基因组DNA提取及RAPD引物筛选

以胡卢巴叶片为材料,利用改良CTAB法提取基因组DNA并进行质量检测和含量测定,在优化RAPD反应体系的基础上,对100条RAPD随机引物进行了筛选.结果表明,采用改良CTAB法能从胡卢巴叶片中提取到高质量的DNA,可以用于RAPD分析;从100条随机引物中筛选出了16条显示胡卢巴遗传差异的多态性引物,为胡卢巴遗传多样性研究提供分子依据.     

Comparison of Molecular Biological Techniques (RAPD, ARDRA and SSrRNA Gene Sequencing) on the Phylogenetic Relationships of Lower Taxa within Spirotrichous Ciliates

The evolutionary relationships of seven spirotrichous ciliates (3 stichotrichs: Oxytricha saltans, O. Ferruginea, Stylonychia mytilus; 4 hypotrichs: Uronychia transfuga, Diophrys appendiculata, Aspidisca steini, Euplotes vannus) inferred from the SSrRNA (small subunit rRNA) gene sequences and the polymorphic patterns of RAPD (random amplified polymorphic DNA) and ARDRA (amplified ribosomal DNA restriction analyses) fingerprinting are constructed. Compared with that of morphometric characters, the dendrograms from SSrRNA gene using three different calculation methods (distance matrix, maximum-parsimony, and UPGMA) agree with the morphological division into two clades, Oxytricha - Stylonychia and Uronychia - Diophrys - Aspidisca - Euplotes, though the branching orders within the hypotrichous ciliates are slightly different from morphometric analyses.     

高空气球搭载诱导水稻育性降低突变体RAPD分析

选用300对引物，对经高空气球30km搭载飞行8h后返地种植筛选获得的水稻育性降低突变体基因组进行DNA多态性分析，结果发现：育性降低突变体与原种之间存在多态性．     

Rapid establishment of pure lines of silver carp (Hypophthalmichthys molirix) and bighead carp (Aristichthys nobilis)

The diversity of gynogenetic, artificial sex reversal and natural silver carp and bighead carp is examined using randomly amplified polymorphic DNA (RAPD) method.All of the 187 bands are obtained and 19 (10.16%) of them are polymorphic in gynogenetic silver carp.Meanwhile 32 (15.61%) out of 205 bands are polymorphic in control group.In gynogenetic bighead carp a total of 232 bands are identified and 11 (4.74%) out of them are polymorphic, while 25 (10.37%) out of 241 bands are polymorphic in control group.The genetic distance of four populations is calculated and it is 0.102 and 0.023 for gynogenetic silver carp and gynogenetic bighead carp respectively.The values of natural silver carp and bighead carp are 0.161 and 0.104.From the UPGMA trees constructed based on genetic distance, the sex reversal individuals that match with the gynogenetic female individuals are picked out.A new breeding process of establishing a pure line is developed.     

乌头基因组DNA提取与RAPD反应体系优化

采用改进方法提取乌头基因组DNA,建立乌头RAPD分析的PCR反应体系,成功地进行了RAPD扩增,筛选出乌头RAPD的最佳方案为:25μl反应体系中包括dNTPs 0.06 mmol·L-1,引物0.2μmol.μL-1,模板DNA40 ng,Taq酶1U,Mg2+ 4 mmol·L-1,10×buffer缓冲液2.5μl,其余部分为无菌超纯水。PCR扩增循环为95℃3 min,38 ℃1 min,72 ℃2min 1次循环;94 ℃=1 min,38℃1 min,72 ℃2 min,45次循环,最后72℃延伸10 min。     

中国栗属特有种保守RAPD片段分析

利用RAPDDNA分子标记技术研究了栗属特有种(板栗、茅栗和锥栗)特异保守片段。结果表明，从520多个随机引物中筛选出有扩增产物，且多态性丰富的10个引物，其中引物B08(GTCCACACGG)分别在所有供试的板栗、茅栗和锥栗中均扩增出各自特异的DNA片段，其片段大小分别为2050，900和1500bp。这些DNA片段在种水平上的高度保守性，可以用来区分这3个种。此次研究结果同时也说明RAPD标记是鉴别近缘种的理想工具。     

斜带石斑鱼、赤点石斑鱼RAPD和线粒体Cyt b基因序列变异分析

斜带石斑鱼和赤点石斑鱼为具有重要经济价值的海水鱼类。对这两种石斑鱼进行了随机扩增多态DNA(RAPD)和线粒体细胞色素b(Cyt b)基因序列变异分析。计算多态位点百分率、基因多样性和香农信息指数等遗传参数,以此评估种内遗传变异水平;通过统计变异位点、平均核苷酸差异数、核苷酸多样性以及种间平均每位点核苷酸替代数进行基因序列变异分析,并构建UPGMA系统树。结果表明:斜带石斑鱼的遗传多样性水平高于赤点石斑鱼,这可能与它们在种内特定遗传结构、分布范围大小、自然资源状况的差异有关;斜带石斑鱼和赤点石斑鱼之间检测到的种特异RAPD条带以及基因序列的变异,可作为种间分子鉴定标记。     

利用RAPD研究星天牛地理种群间亲缘关系

利用随机扩增的多态性DNA(RAPD)方法对分布在我国甘肃、宁夏、内蒙、河北、西安、青岛、福建、江苏等16个地区,美国纽约、芝加哥、新泽西3个地区和韩国2个地区的光肩星天牛及其近缘种星天牛共23个地理种群进行了研究.实验筛选出17个多态性引物产生了219个多态性位点,RAPD聚类分析表明,美国和中国的光肩星天牛种群之间存在显著差异,遗传关系较远.